In VitroAntiparasitic and Apoptotic Effects of Antimony Sulfide Nanoparticles onLeishmania infantum

Visceral leishmaniasis is one of the most important sever diseases in tropical and subtropical countries. In the present study the effects of antimony sulfide nanoparticles onLeishmania infantum in vitrowere evaluated. Antimony sulfide NPs (Sb2S5) were synthesized by biological method fromSerratia marcescensbacteria. Then the cytotoxicity effects of different concentrations (5, 10, 25, 50, and 100 μg/mL) of this nanoparticle were assessed on promastigote and amastigote stages ofL. infantum. MTT method was used for verification results of promastigote assay. Finally, the percentages of apoptotic, necrotic, and viable cells were determined by flow cytometry. The results indicated the positive effectiveness of antimony sulfide NPs on proliferation of promastigote form. The IC50(50% inhibitory concentration) of antimony sulfide NPs on promastigotes was calculated 50 μg/mL. The cytotoxicity effect was dose-dependent means by increasing the concentration of antimony sulfide NPs, the cytotoxicity curve was raised and the viability curve of the parasite dropped simultaneously. Moreover, the IC50of antimony sulfide NPs on amastigote stage was calculated 25 μg/mL. On the other hand, however, antimony sulfide NPs have a low cytotoxicity effect on uninfected macrophages but it can induce apoptosis in promastigote stage at 3 of 4 concentrations.

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Kaempferol Induces DNA Damage and Inhibits DNA Repair Associated Protein Expressions in Human Promyelocytic Leukemia HL-60 Cells

The American Journal of Chinese Medicine ◽

10.1142/s0192415x1550024x ◽

2015 ◽

Vol 43 (02) ◽

pp. 365-382 ◽

Cited By ~ 20

Author(s):

Lung-Yuan Wu ◽

Hsu-Feng Lu ◽

Yu-Cheng Chou ◽

Yung-Luen Shih ◽

Da-Tian Bau ◽

...

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Protein Expression ◽

Ataxia Telangiectasia ◽

Repair System ◽

Dapi Staining ◽

Viable Cells ◽

Protein Expressions ◽

Dose Dependent

Numerous evidences have shown that plant flavonoids (naturally occurring substances) have been reported to have chemopreventive activities and protect against experimental carcinogenesis. Kaempferol, one of the flavonoids, is widely distributed in fruits and vegetables, and may have cancer chemopreventive properties. However, the precise underlying mechanism regarding induced DNA damage and suppressed DNA repair system are poorly understood. In this study, we investigated whether kaempferol induced DNA damage and affected DNA repair associated protein expression in human leukemia HL-60 cells in vitro. Percentages of viable cells were measured via a flow cytometry assay. DNA damage was examined by Comet assay and DAPI staining. DNA fragmentation (ladder) was examined by DNA gel electrophoresis. The changes of protein levels associated with DNA repair were examined by Western blotting. Results showed that kaempferol dose-dependently decreased the viable cells. Comet assay indicated that kaempferol induced DNA damage (Comet tail) in a dose-dependent manner and DAPI staining also showed increased doses of kaempferol which led to increased DNA condensation, these effects are all of dose-dependent manners. Western blotting indicated that kaempferol-decreased protein expression associated with DNA repair system, such as phosphate-ataxia-telangiectasia mutated (p-ATM), phosphate-ataxia-telangiectasia and Rad3-related (p-ATR), 14-3-3 proteins sigma (14-3-3σ), DNA-dependent serine/threonine protein kinase (DNA-PK), O6-methylguanine-DNA methyltransferase (MGMT), p53 and MDC1 protein expressions, but increased the protein expression of p-p53 and p-H2AX. Protein translocation was examined by confocal laser microscopy, and we found that kaempferol increased the levels of p-H2AX and p-p53 in HL-60 cells. Taken together, in the present study, we found that kaempferol induced DNA damage and suppressed DNA repair and inhibited DNA repair associated protein expression in HL-60 cells, which may be the factors for kaempferol induced cell death in vitro.

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Silymarin: Friend or Foe of UV Exposed Keratinocytes?

Molecules ◽

10.3390/molecules24091652 ◽

2019 ◽

Vol 24 (9) ◽

pp. 1652 ◽

Cited By ~ 3

Author(s):

Fidrus ◽

Ujhelyi ◽

Fehér ◽

Hegedűs ◽

Janka ◽

...

Keyword(s):

Pyrimidine Dimer ◽

Ros Generation ◽

Keratinocyte Cell ◽

Pre Treatment ◽

Different Origins ◽

Apoptotic Effects ◽

Dose Dependent ◽

Higher Doses

The application of natural plant extracts in UV-protection is popular and intensively studied. Silymarin (from Silibum marianum), a naturally occurring polyphenol, has recently received attention due to its antioxidant, anti-inflammatory and anti-apoptotic effects. However, its role in the UV-mediated keratinocyte cell response is still controversial. In this study, we investigated the effects of Silibum marianum extracts with different origins and formulations on UVA-exposed HaCaT keratinocytes in vitro. Our results show, that silymarin treatment caused an inverse dose-dependent photosensitivity relationship (at higher doses, a decrease in cell viability and ROS production) after UVA exposure. The attenuation of the UVA-induced ROS generation after silymarin treatment was also observed. Moreover, silymarin pre-treatment increased the cyclobutane pyrimidine dimer photolesions in keratinocytes after UVA exposure. These results indicated the dual role of silymarin in UVA-exposed keratinocytes. It scavenges ROS but still induces phototoxicity. Based on our results dermatological applications of silymarin and related compounds should be considered very carefully.

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Synthesis, Characterization, and Interaction with Biomolecules of Platinum(II) Complexes with Shikimic Acid-Based Ligands

Bioinorganic Chemistry and Applications ◽

10.1155/2013/565032 ◽

2013 ◽

Vol 2013 ◽

pp. 1-12 ◽

Cited By ~ 4

Author(s):

Yan Peng ◽

Min-Min Zhang ◽

Zhen-Feng Chen ◽

Kun Hu ◽

Yan-Cheng Liu ◽

...

Keyword(s):

Shikimic Acid ◽

Covalent Binding ◽

Carbon Chain ◽

Water Soluble ◽

Cytotoxic Activities ◽

Experimental Conditions ◽

H Nmr ◽

Mtt Method ◽

Low Cytotoxicity

Starting from the active ingredient shikimic acid (SA) of traditional Chinese medicine and NH2(CH2)nOH, (n=2–6), we have synthesized a series of new water-soluble Pt(II) complexes PtLa–eCl2, where La–eare chelating diamine ligands with carbon chain covalently attached to SA (La–e= SA-NH(CH2)nNHCH2CH2NH2; La,n=2; Lb,n=3; Lc,n=4; Ld,n=5; Le,n=6). The results of the elemental analysis, LC-MS, capillary electrophoresis, and1H,13C NMR indicated that there was only one product (isomer) formed under the present experimental conditions, in which the coordinate mode of PtLa–eCl2was two-amine bidentate. Theirin vitrocytotoxic activities were evaluated by MTT method, where these compounds only exhibited low cytotoxicity towards BEL7404, which should correlate their low lipophilicity. The interactions of the five Pt(II) complexes with DNA were investigated by agarose gel electrophoresis, which suggests that the Pt(II) complexes could induce DNA alteration. We also studied the interactions of the Pt(II) complexes with5′-GMP with ESI-MS and1H NMR and found that PtLbCl2, PtLcCl2, and PtLdCl2could react with5′-GMP to form mono-GMP and bis-GMP adducts. Furthermore, the cell-cycle analysis revealed that PtLbCl2, PtLcCl2cause cell G2-phase arrest after incubation for 72 h. Overall, these water-soluble Pt(II) complexes interact with DNA mainly through covalent binding, which blocks the DNA synthesis and replication and thus induces cytotoxicity that weakens as the length of carbon chain increases.

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Effect of Clinoptilolite and Sepiolite Nanoclays on Human and Parasitic Highly Phagocytic Cells

BioMed Research International ◽

10.1155/2015/164980 ◽

2015 ◽

Vol 2015 ◽

pp. 1-12 ◽

Cited By ~ 8

Author(s):

Yanis Toledano-Magaña ◽

Leticia Flores-Santos ◽

Georgina Montes de Oca ◽

Alfonso González-Montiel ◽

Juan-Pedro Laclette ◽

...

Keyword(s):

Cell Line ◽

Human Peripheral Blood ◽

Dependent Manner ◽

Blood Monocytes ◽

Phagocytic Cells ◽

Need To Evaluate ◽

Potential Applications ◽

Low Cytotoxicity ◽

Dose Dependent

Nanoclays have potential applications in biomedicine raising the need to evaluate their toxicity inin vitromodels as a first approach to its biocompatibility. In this study,in vitrotoxicity of clinoptilolite and sepiolite nanoclays (NC) was analyzed in highly phagocytic cultures of amoebas and human and mice macrophages. While amebic viability was significantly affected only by sepiolite NC at concentrations higher than 0.1 mg/mL, the effect on macrophage cultures was dependent on the origin of the cells. Macrophages derived from human peripheral blood monocytes were less affected in viability (25% decrease at 48 h), followed by the RAW 264.7 cell line (40%), and finally, macrophages derived from mice bone marrow monocytes (98%). Moreover, the cell line and mice macrophages die mainly by necrosis, whereas human macrophages exhibit increased apoptosis. Cytokine expression analysis in media of sepiolite NC treated cultures showed a proinflammatory profile (INFγ, IL-1α, IL-8, and IL-6), in contrast with clinoptilolite NC that induced lees cytokines with concomitant production of IL-10. The results show that sepiolite NC is more toxic to amoebas and macrophages than clinoptilolite NC, mostly in a time and dose-dependent manner. However, the effect of sepiolite NC was comparable with talc powder suggesting that both NC have low cytotoxicityin vitro.

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Antiviral effect of copper chloride on feline calicivirus and Synergy with Ribavirin in vitro

10.21203/rs.2.20385/v2 ◽

2020 ◽

Author(s):

Dengliang Li ◽

Zhanding Cui ◽

Guohua Li ◽

Liangting Zhang ◽

Ying Zhang ◽

...

Keyword(s):

Copper Chloride ◽

Antiviral Effect ◽

Antagonistic Effect ◽

Drug Candidate ◽

Feline Calicivirus ◽

Antiviral Effects ◽

Low Cytotoxicity ◽

Effect Of Copper ◽

Dose Dependent

Abstract Background: Feline calicivirus (FCV) is a common pathogen causing widely prevalent upper respiratory disease for kitten and felines in recent years. Due to the substantial genetic variability of the viral genes, existing vaccines cannot provide complete protection. Therefore, researches on FCV antiviral drugs have received much attention. Results: In this study, we found that copper chloride had dose-dependent antiviral effects against FCV in F81 cells. We also found that the combination of copper chloride and ribavirin had a synergistic effect against FCV in the F81 cells. In contrast, the combination of and horse anti-FCV immunoglobulin F(ab’) 2 showed an antagonistic effect, likely because the copper chloride has an effect on the F(ab’) 2 immunoglobulin; however, further research is needed to clarify this supposition. Conclusions: In summary, we found that copper chloride had low cytotoxicity and significant antiviral effects against FCV in F81 cells, providing a new drug candidate for the prevention and treatment of FCV infection.

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Antiviral effect of copper chloride on feline calicivirus and synergy with ribavirin in vitro

10.21203/rs.2.20385/v3 ◽

2020 ◽

Author(s):

dengliang li ◽

Zhanding Cui ◽

Guohua Li ◽

Liangting Zhang ◽

Ying Zhang ◽

...

Keyword(s):

Copper Chloride ◽

Antiviral Effect ◽

Antagonistic Effect ◽

Drug Candidate ◽

Feline Calicivirus ◽

Antiviral Effects ◽

Low Cytotoxicity ◽

Effect Of Copper ◽

Dose Dependent

Abstract Background: Feline calicivirus (FCV) is a common and highly prevalent pathogen causing upper respiratory diseases in kittens and felines in recent years. Due to the substantial genetic variability of the viral genes, existing vaccines cannot provide complete protection. Therefore, research on FCV antiviral drugs has received much attention.Results: In this study, we found that copper chloride had dose-dependent antiviral effects on FCV in F81 cells. We also found that the combination of copper chloride and ribavirin had a synergistic protective effect against FCV in F81 cells. In contrast, the combination of copper chloride and horse anti-FCV immunoglobulin F(ab')2 showed an antagonistic effect, likely because copper chloride has an effect on F(ab')2 immunoglobulin; however, further research is needed to clarify this supposition.Conclusions: In summary, we found that copper chloride had low cytotoxicity and significant antiviral effects on FCV in F81 cells, providing a new drug candidate for the prevention and treatment of FCV infection.

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Sustained-Release and pH-Adjusted Alginate Microspheres-Encapsulated Doxorubicin Inhibit the Viabilities in Hepatocellular Carcinoma-Derived Cells

Pharmaceutics ◽

10.3390/pharmaceutics13091417 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1417

Author(s):

Cheng-Tang Pan ◽

Ruei-Siang Yu ◽

Chih-Jung Yang ◽

Lih-Ren Chen ◽

Zhi-Hong Wen ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Sustained Release ◽

Flow Rate ◽

Calcium Alginate ◽

Ultrasonic Atomization ◽

Median Diameter ◽

Cytotoxicity Effects ◽

Micrometer Scale ◽

Dose Dependent

The objective of this study aimed to develop biodegradable calcium alginate microspheres carrying doxorubicin (Dox) at the micrometer-scale for sustained release and the capacity of pH regulatory for transarterial chemoembolization. Ultrasonic atomization and CaCl2 cross-linking technologies were used to prepare the microspheres. A 4-by-5 experiment was first designed to identify imperative parameters. The concentration of CaCl2 and the flow rate of the pump were found to be critical to generate microspheres with a constant volume median diameter (~39 μm) across five groups with different alginate: NaHCO3 ratios using each corresponding flow rate. In each group, the encapsulation efficiency was positively correlated to the Dox-loading %. Fourier-transform infrared spectroscopy showed that NaHCO3 and Dox were step-by-step incorporated into the calcium alginate microspheres successfully. Microspheres containing alginate: NaHCO3 = 1 exhibited rough and porous surfaces, high Young’s modulus, and hardness. In each group with the same alginate: NaHCO3 ratio, the swelling rates of microspheres were higher in PBS containing 10% FBS compared to those in PBS alone. Microspheres with relatively high NaHCO3 concentrations in PBS containing 10% FBS maintained better physiological pH and higher accumulated Dox release ratios. In two distinct hepatocellular carcinoma-derived cell lines, treatments with microspheres carrying Dox demonstrated that the cell viabilities decreased in groups with relatively high NaHCO3 ratios in time- and dose-dependent manners. Our results suggested that biodegradable alginate microspheres containing relatively high NaHCO3 concentrations improved the cytotoxicity effects in vitro.

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Small Molecule Factor D Inhibitors Block Complement Activation in Paroxysmal Nocturnal Hemoglobinuria and Atypical Hemolytic Uremic Syndrome

Blood ◽

10.1182/blood.v126.23.275.275 ◽

2015 ◽

Vol 126 (23) ◽

pp. 275-275 ◽

Cited By ~ 4

Author(s):

Eleni Gavriilaki ◽

Jane A Thanassi ◽

Guangwei Yang ◽

Xuan Yuan ◽

Mngjun Huang ◽

...

Keyword(s):

Atypical Hemolytic Uremic Syndrome ◽

Inhibitory Effect ◽

Cell Killing ◽

Dependent Manner ◽

Hemolytic Assay ◽

Complement Inhibitors ◽

Viable Cells ◽

Uremic Syndrome ◽

Dose Dependent

Abstract Introduction: Factor D is a highly specific serine protease that cleaves factor B as its only substrate. It is the rate-limiting step of the alternative pathway of complement (APC). Therefore, factor D is a promising therapeutic target in diseases of excess activation of the APC, such as paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS). Terminal complement inhibition by eculizumab is currently the treatment of choice for PNH and aHUS. Eculizumab must be administered intravenously and indefinitely in PNH; moreover, up to 20% of PNH patients treated with eculizumab have symptomatic hemolysis due to extravascular hemolysis. Glycosylphosphatidylinositol (GPI) anchor protein deficient erythrocytes from PNH patients or erythrocytes from animals can be used to test novel complement inhibitors for PNH; however, there are no in vitro assays available to model complement-mediated attack in aHUS. Here, we demonstrate that the modified Ham test (PIGA-null reagent cell line exposed to serum from aHUS patients) is the first reliable in vitro model to test complement inhibition for aHUS. In addition, we demonstrate that small molecule factor D inhibitors specifically block the APC in PNH and aHUS. Method: Three factor D inhibitors (ACH-3856, ACH-4100, ACH-4471) that reversibly bind factor D and are being developed for oral administration were studied. We obtained blood samples from 5 PNH and 4 aHUS patients after written informed consent. Serum and erythrocytes from PNH patients on eculizumab were collected within 60 minutes of eculizumab administration. All 3 compounds were tested in half-log dilutions ranging from 0.001 ìM to 10 ìÌ. The binding affinity to factor D was determined by surface plasmon resonance (Biacore) analysis. The inhibitory effect on factor D protease was evaluated biochemically using a natural substrate consisting of C3b and the complement factor B. The inhibition of APC-mediated hemolysis was first evaluated with rabbit erythrocytes incubated with 8% normal human serum (NHS). The inhibition of APC-mediated C3 fragment deposition was also initially evaluated with rabbit erythrocytes incubated with 20% C5-depleted NHS (to mimic the effect of eculizumab) by flow cytometry. To confirm the observation with rabbit erythrocytes, erythrocytes from PNH patients were also tested in the hemolytic assay (20% acidified NHS). Regarding aHUS, the inhibitory effect of the compounds was evaluated in the recently described modified Ham test. Briefly, PIGA-null TF-1 cells were incubated with 20% of aHUS serum for 30 minutes at 37 o C and complement-mediated cell killing was evaluated using the cell proliferation reagent (WST-1). Absorbance was measured and expressed as a ratio to the heat-inactivated control (% non-viable cells) Results: All 3 small molecules showed high binding affinity to human factor D and effectively inhibited factor D proteolytic activity in a dose-dependent manner (mean IC50 9.8-20 nM). Similarly, we observed a dose-dependent inhibition of hemolysis on rabbit erythrocytes (EC50 9-17 nM). C3 fragment deposition on rabbit erythrocytes after incubation with C5-depleted serum was reduced to undetectable levels with all 3 compounds from concentrations of 0.1 ìÌ and above. Next, we studied samples from 5 PNH patients (2 treatment naïve and 3 on eculizumab) and 4 aHUS patients (1 on eculizumab, 1 in acute phase and on eculizumab and 2 in remission). When tested on the hemolytic assay with erythrocytes from PNH patients and acidified NHS, factor D inhibitors significantly reduced hemolysis in a dose-dependent manner from concentrations as low as 0.01 ìM. Lastly, cell killing was observed as previously reported in the presence of the serum from aHUS patients in the modified Ham test and addition of factor D inhibitors caused significant dose-dependent reduction in the cell killing at concentration as low as 0.01 ìÌ. Mean percentage of non-viable cells before and after addition of compound ACH-4471 is shown in Figure 1. Conclusions: We demonstrate that factor D inhibitors, potentially the first-ever oral complement inhibitors, efficiently block hemolysis of PNH cells in vitro and mitigate the accumulation of C3 fragments. Importantly, our findings indicate that the recently described modified Ham test can be used as a preclinical model to test complement inhibitors in aHUS. Figure 1. Figure 1. Disclosures Thanassi: Achillion Pharmaceuticals: Employment. Yang:Achillion Pharmaceuticals: Employment. Huang:Achillion Pharmaceuticals: Employment. Brodsky:Alexion Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees.

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The Effects of Thymoquinone on Viability, and Anti-apoptotic Factors (BCL-XL, BCL-2, MCL-1) in Prostate Cancer (PC3) Cells: An In Vitro and Computer-Simulated Environment Study

Advanced Pharmaceutical Bulletin ◽

10.15171/apb.2019.058 ◽

2019 ◽

Vol 9 (3) ◽

pp. 490-496 ◽

Cited By ~ 1

Author(s):

Javad Saffari_Chaleshtori ◽

Ehsan Heidari-Sureshjani ◽

Fahimeh Moradi ◽

Esfandiar Heidarian

Keyword(s):

Root Mean Square ◽

Radius Of Gyration ◽

Mean Square ◽

Simulated Environment ◽

Environment Study ◽

Pc3 Cells ◽

Cytotoxicity Effects ◽

Apoptotic Effects ◽

Apoptotic Factors

Purpose: Since active plant ingredients can induce apoptosis in many tumors, in this study we evaluate the apoptotic effects of thymoquinone (TQ) on PC3 cells. Also, we predicted the interaction of TQ with BCL-XL, BCL-2, and MCL-1anti-apoptotic factors by computer-simulated environment. Methods: PC3 cells were treated with different concentrations of TQ (0- 80 µM) and IC50 was determined using 3-(4, 5-dimethylthiaztol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Apoptotic and cytotoxicity effects of TQ were analyzed using flowcytometry and comet assay, respectively. Changes in energy and the molecular interactions of TQ with BCL-XL, BCL-2 and MCL-1 anti-apoptotic factors were investigated using simulation. Results: IC50 value was 40 µM. TQ led to the destruction of the genome of PC3 cells and inducing apoptosis. Molecular dynamics (MD) revealed that the root mean-square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (Rg), and the number of hydrogen and hydrophobic bonds between TQ and residues of BCL-2, BCL-XL and MCL-1were significantly (P<0.001) changed. TQ makes a more stable and stronger connection with BCL-XL compared to BCL-2 and MCL-1 and inhibits BCL-XL non-competitively. Conclusion: Our results demonstrated that TQ not only led to apoptosis, at least partly, due to reduction in the Coil, Turn, and Bend structure of BCL-XL but also caused a decrease in the Rg and RMSD value of BCL-XL, MCL-1, and BCL-2.

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Antiviral effect of copper chloride on feline calicivirus and Synergy with Ribavirin in vitro

10.21203/rs.2.20385/v1 ◽

2020 ◽

Author(s):

dengliang li ◽

Zhanding Cui ◽

Guohua Li ◽

Liangting Zhang ◽

Ying Zhang ◽

...

Keyword(s):

Copper Chloride ◽

Antiviral Effect ◽

Antagonistic Effect ◽

Drug Candidate ◽

Feline Calicivirus ◽

Antiviral Effects ◽

Low Cytotoxicity ◽

Effect Of Copper ◽

Dose Dependent

Abstract Background: Feline calicivirus (FCV) is a common pathogen causing widely prevalent upper respiratory disease for kitten and felines in recent years. Due to the substantial genetic variability of the viral genes, existing vaccines cannot provide complete protection. Therefore, researches on FCV antiviral drugs have received much attention. Results: In this study, we found that copper chloride had dose-dependent antiviral effects against FCV in F81 cells. We also found that the combination of copper chloride and ribavirin had a synergistic effect against FCV in the F81 cells. In contrast, the combination of and horse anti-FCV immunoglobulin F(ab’)2 showed an antagonistic effect, likely because the copper chloride has an effect on the F(ab’)2 immunoglobulin; however, further research is needed to clarify this supposition.Conclusions: In summary, we found that copper chloride had low cytotoxicity and significant antiviral effects against FCV in F81 cells, providing a new drug candidate for the prevention and treatment of FCV infection.

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